ウシ発情周期中に発育する第1卵胞波主席卵胞の生理学的 ......Title ウシ発情周期中に発育する第1卵胞波主席卵胞の生理学的性質 ― 発育動態、機能および受胎性に及ぼす影響

Titleウシ発情周期中に発育する第１卵胞波主席卵胞の生理学的性質　― 発育動態、機能および受胎性に及ぼす影響 ―( 本文(Fulltext) )

Author(s) 三浦, 亮太朗

Report No.(DoctoralDegree) 博士(獣医学) 甲第440号

Issue Date 2015-03-13

Type 博士論文

Version ETD

URL http://hdl.handle.net/20.500.12099/51006

※この資料の著作権は、各資料の著者・学協会・出版社等に帰属します。

1

2014

i

1

1 W1DF

9

2 1. 10

2 2. 1 7 W1DF W2DF

10

2 3. 2 W1DF W2DF

E2 14

2 4. 3 W1DF W2DF

16

2 5. 4 W1DF W2DF

P4 21

2 6. 23

2 7. 26

2 8. 32

33

ii

1 W1DF 57

3 1. 58

3 2. 1 7 W1DF P4

W1DF W1DF

58

3 3. 2 W1DF 5

W1DF P4

60

3 4. 63

3 5. 64

3 6. 66

68

1 W1DF

77

4 1. 78

4 2. 1 W1DF W1DF

P4 78

iii

4 3. 2 W1DF

80

4 4. 82

4 5. 84

4 6. 87

88

92

100

101

iv

P4 Progesterone

E2 Estradiol-17

A4 Androstendion

GnRH Gonadotropin releasing hormone

FSH Follicle stimulating hormone

LH Luteinizing hormone

W1DF The first follicular wave dominant follicle 1

W2DF The second follicular wave dominant follicle 2

PG Prostaglandin

BFA Blood flow area

BF% The percentage of follicle circumference with blood flow signals

EIA Enzyme immuno assay

BSA Bovine serum albumin

cDNA complementary DNA DNA

PCR Polymerase chain reaction

LHr LH receptor LH

CYP19A1 Aromatase

COX-2 Cyclooxygenase-2

NO Nitric oxide

iNOS Inducible nitric oxide synthase

eNOS Endotherial nitric oxide synthase

VEGF Vascular endotherial growth factor

FGF Fibroblast growth factor

v

StAR Steroidogenic acute regulatory protein

P450-scc Cytochrome P450 side chain cleavage enzyme P450

3 -HSD 3 -hydroxysteroid dehydrogenase 3 -

GAPDH Glyceraldehyde-3-phosphate dehydrogenase 3

W1CL Corpus luteum formed after ovulation of W1DF W1DF

W2CL Corpus luteum formed after ovulation of W2DF W2DF

TMR Total mixed ration

BCS Body condition score

1

2

9,000

Bos primigenius

FAO

2007 13 6,000

31 29 19

10 8 3

2 1 8,000 9,700 8,200

5,100 440

1

1 1960 2.0

1980 18.1 1990 32.5 2000 52.5 2012

72.1 50

1951 1996 5,000 kg

3

9,000 kg 18

1975 5,826 kg 1980 6,339 kg 1990

7,798 kg 2000 8,794 kg 2010 9,286 kg

24

1 1960 1.2 1980

5.9 1990 11.6 2000 24.2 2012 41.8

72 92

95 44

100

55

72

32 50 60 77 90

4

6 12

30 1

21 30

18 24 30 14 18

4 6 2 30

30

E2 30 E2

GnRH

LH 80 94

LH

8 7

PG F2 54

11 51 67 P4 68

P4

P4 P4 57

2 3

5

1 2

41

3 4mm

8 41 1 41

1

1 1 W1DF

W1DF 8 11 P4

W1DF

41 2

2 3

7 16 17 19 28 39

69 74 88 108 3 66 76

10 W1DF

48

2 2 9 11

3 8 9 4 3

3 15 16 4 2 3

2 2 vs. 3 16 vs. 19

2 vs. 3 19 20 vs. 22 23 4

2 3 1

1 1

6

GnRH FSH LH

80 GnRH 65

FSH LH FSH

6 LH

GnRH 107 LH

36

E2 P4

P4 LH P4

LH 15 52 LH

P4

P4 LH

E2 E2 E2

GnRH LH

81

63

49 FSH LH

1

2 W2DF

7

FSH

FSH 62

8 41 3 4mm

17 34

1 1.5 2.0 mm

37 2 3

LH E2 13 E2 FSH

38

FSH

LH FSH LH

LH

43

8.5mm 7.7mm

35 W1DF P4

W2DF

P4 LH

E2 LH

W2DF

1

W1DF

1

W1DF P4 2

W2DF P4

LH

8

W1DF W2DF P4

LH

W1DF W2DF

W1DF

W1DF W2DF

W1DF 1

W1DF

P4 20

E2 99

W1DF

2 W1DF

W2DF

3 W1DF W1DF

P4

4 W1DF

9

1 W1DF

10

2 1

1

W1DF 8 10

41

W1DF

9 53 93 103 W1DF

2 9 10

88 1

W1DF P4

2 W2DF

P4 P4

W1DF

W2DF

2 2 1 7 W1DF W2DF

2 2 1

11

8

5.0 1.8

TMR

2 2 2

8 1 n 4 2 n 4

2

8 n 8

1 12

Day 0

Day 3 5

Day 6

Day 0 6 1 1

2 Day 7 2

Day 2 W1DF GnRH 100 μg

GnRH 36 W1DF

Day 0 1

Day 3 5

Day 6

12

Day 0 6 1 1

1 Fig. 2.1.

2 2 3

Day 3 5 5.0MHz

HS 101V

Day 6 8.0MHz 1739

RS GE

LOGIQ Book XP GE

BFA BF%

BFA Photoshop 5.5 Adobe Systems

Inc. San Jose CA USA Pop Imaging

BFA

Fig. 2.2.

BF

Photoshop 5.5

13

BF

2 2 4

Day 0 6 21G

TERUMO 10ml

4 2,000 g 20 P4 30

2 2 5 P4

P4 2 EIA EIA

64 P4 200μl

1.0ml Mix-EVR

30 40 30

24 5ml

40

50

7.12g NA2HPO4 8.5g NaCl 1.0g BSA pH 7.2 1N-HCL

200μl 20 P4 84

EIA IgG 12.5μg/ml

96 ELISA Corning Glass Works Corning New York USA

15μl 100μl P4 OK-1

1 300,000 100μl horseradish peroxidase 1 50,000 4 20

14

P4 0.05 50ng/ml

5.2 7.4%

2 2 6

P4

2

Bartlet Post-hoc Schefe

Day 6 F

Student t 10 P < 0.1

5 P < 0.05 StatView

Abacus Concepts Inc. Berkeley CA USA

2 3 2 W1DF W2DF

E2

2 3 1

2 2

2 3 2

1 1 2

Day 6 PGF2α 25 mg

F

48 Day 8 GnRH Day 6 8

15

9

Day 6 9 1 1 2

Fig. 2.5.

2 3 3

Day 6 8 9 8.0MHz

1739 RS GE

LOGIQ Book XP GE

1 BFA BF%

2 3 4

Day 6 9 21G

TERUMO 10ml

4 2,000 g 20 P4 E2

30

2 3 5 P4 E2

P4 1 EIA P4

80 4.7 6.5

E2 Day 8 E2 2.0ml

6.0ml DM-301

16

60 60 30

24 10ml

40

50

200μl 20 E2 10

E2 80

15μl 100μl E2 AS-A 1 20,000 4

24 100μl horseradish peroxidase 1 150,000 4

2 E2 2 2,000 pg/ml

6.5 7.6%

2 3 6

P4

2

Bartlet

Post-hoc Schefe Day 8 E2

F Student t P < 0.1

P < 0.05 StatView Abacus

Concepts Inc. Berkeley CA USA

2 4 3 W1DF W2DF

17

2 4 1

7

4.0 1.1 )

TMR

2 4 2

2 LH PGF2α

40 Day 8 LH GnRH 24 Day 9

4 LH 1 n 7 LH

2 n 7 LH 1 n 6 LH

2 n 6 7

1

6

3 Fig. 2.9.

2 4 3

HS-1500

7.5MHz

HCV-3710MV

18

40cm A-

18G

5.0ml

TriPure Isolation Reagent Roche

Diagnostics GmbH Mannheim Germany 1.0ml 80 mRNA

4 500 g 10

EIA P4 E2 A4 30

2 4 4 P4 E2 A4

40μl

1.0ml Mix-EVR

30 40 30

24 5ml

40

50

200μl 20

5 P4 E2 A4 82 81

85 P4 E2 2 2 5. 2 3 5.

19

P4 5.5% 7.4% E2

6.2 8.6% A4

15μl 100μl A4 A-3-CMO-BSA

1 750,000 100μl A-3-CMO-horseradish peroxidase

1 500,000 4 24 A4 7.8 8,000 pg/ml

4.3 5.2%

2 4 5 RNA PCR

Total RNA 200μl

15 3 4 36,000 g 15

TriPure Isolation Reagent

1.5ml 500μl

3-Propanol 15 10 4 36,000 g

15 RNA

500μl 80 4 36,000 g 5

5 6μl

DEPC

Superscript VIRO Reaction Mix Invitrogen. Carlsbad CA USA

Nano-Drop 2000 Thermo scientific Wilmington DE USA RNA

RQ1 RNase-Free DNase (Promega Madison WI USA) DNase

RNA 1μg RNA 1μl DNase 10×Reaction Buffer 2μl

RNase-Free DNase 37 30 1μl DNase Stop

Solution 65 10 DNase 5×VILO

20

Reaction Mix 4μl 10×Super Script Enzyme 2μl 20μl DEPC

25 10 42 60 85 5 cDNA

2 4 6 PCR mRNA

PCR Tli RNaseH Plus Roter-Gene Q

7.5μl SYBR Green Master Mix 5.75μl DEPC

Forward Reverse 0.375μl 1.0μl

mRNA LH LHr CYP19A1

2 COX-2 PGE2 PGES PGF2α PGFS

iNOS eNOS

VEGF 120 VEGF164 FGF -2 Steroidogenic acute regulatory

protein StAR P450 cytochrome P450 cholesterol side-chain

cleavage enzymes P450-scc 3β- 3β-hydroxy steroid dehydrogenase

3β-HSD 3 glyceraldehyde-3-phosphate

dehydrogenase GAPDH GAPDH

Primer-3 PCR 94

10 94 30 30 72 30

40 mRNA Table

2.3. LH LHr CYP19A1 mRNA

LH COX-2 PGES PGFS iNOS eNOS

VEGF120 VEGF164 FGF-2 StAR P450-scc 3β-HSD mRNA

21

2 4 7

mRNA F

Student t

Mann–Whitney U P < 0.1 P < 0.05

StatView Abacus Concepts Inc.

Berkeley CA USA

2 5 4 W1DF W2DF

P4

2 5 1

6

4.2 2.1 )

TMR

2 5 2

6 W1DF W1CL n 3 W2DF W2CL n 3

2

6 n 6

22

2

W1DF W2DF Day 8 GnRH 24 36

24 36

Day 10 Day 9 Day

12 15 18 21

Day 10 21 1 1 4

Fig. 2.15.

2 5 3

Day 9 Day 12 15 18 21

8.0MHz 1739 RS GE

LOGIQ

Book XP GE

1 BF

2 2×3.14

Photoshop 5.5

Adobe Systems Inc. San Jose CA USA Pop

23

Imaging

2 5 4

Day 10 21 21G

TERUMO 10ml

4 2,000 g 20 P4 30

2 5 5 P4

P4 1

6.4 8.6%

2 5 6

P4

2

Bartlet Post-hoc

Schefe

F Student t P < 0.1

P < 0.05 StatView

Abacus Concepts Inc. Berkeley CA USA

2 6

24

2 6 1 1

P4 Fig. 2.3. W2 W1

Fig. 2.4. W1DF W2DF

Day 6 W1DF Day 3

4 Day 6 W2DF Day 3 Day 6

Day 6 BFA BF Table 2.1. BFA BF% W1DF

2 6 2 2

Fig. 2.6. W1DF W2DF

P4 Fig. 2.7.

Day 6 P4 W2DF W1DF

P4 PGF2

BFA BF% Fig. 2.8. BFA W1DF Day 9

W2DF Day 6 8

BFA W1DF W2DF Day 6 9

W1DF Day 8 9 W2DF

BF W1DF Day 9 W2DF

Day 6 8 BF

25

W1DF Day 6 9 Day 8 9 W2DF

Day 8 E2 Table 2.2. E2 W1DF

2 6 3 3

LH E2 A4 W1DF P4

Table 2.4. LH P4 E2

Table 2.5.

LHr CYP19A1 mRNA Fig. 2.10. LHr W1DF W2DF

CYP19A COX-2 PGES PGFS mRNA

Fig. 2.11 iNOS eNOS mRNA Fig. 2.12.

iNOS eNOS W1DF VEGF120

VEGF164 FGF-2 mRNA Fig. 2.13. W1DF VEGF120

FGF-2 VEGF164 StAR P450-scc 3β-HSD

mRNA Fig. 2.14. W1DF StAR P450-scc

3β-HSD mRNA

2 6 4 4

Day 9 W1DF W2DF BF Table 2.6. W1DF

BF% Fig. 2.16.

Day 15 18 21 W1CL W2CL

26

Fig. 2.17. Day 12 15 W1CL W2CL

P4 Fig .2.18. P4 W1CL W2CL

2 7

1 2 W1DF W2DF

LH E2 3 E2

A4 LHr

mRNA W1DF 4 W1DF

P4

W1DF W2DF

W1DF P4

LH

1 7

W1DF

W1DF W2DF P4 Day 0

Day 6 W1DF

LH

73 P4 P4 LH

P4 LH

15 52 Adams et al. 5

P4 1

27

LHr 13 1 W1DF W2DF

W1DF P4

LH

1 Day 6 W1DF W2DF

95

42

2

W1DF W2DF

W1DF W2DF

2 PGF2α GnRH

W1DF W2DF Day 6

Day 9 W1DF

W1DF GnRH W2DF

Day 9 W1DF W2DF

LH 27 24

LH 19 21

85 LH PG

28

25 LH

GnRH 24

W1DF BF Day 9

W1DF W1DF

LH

GnRH

26

84

87 GnRH

26

86

GnRH

Day 8 E2 W1DF

GnRH E2

14

70 E2

W1DF E2

W2DF

29

3 2 W1DF LH

mRNA

LH

StAR

P450-scc

3β-HSD

P4 22 P4 C17 17α

C17-20 A4 31 59

A4 CYP19A1

E2 12

E2 A4 W1DF CYP19A1 mRNA

E2

A4

9 W1DF E2 W1DF

A4 LH

LHr mRNA W1DF

LH LHr mRNA 56

1 W1DF P4 W1DF

LH W1DF LHr

LH LHr

W1DF LH

30

2 Day 9 W1DF

W1DF LH

LH

E2 P4

89 LH PG

1 3 PG 21 98

26 61

26 PGE2

71

NO

83 104 VEGF FGF

82 105

LH

COX-2 PGES PGFS mRNA

LH

W1DF W2DF

eNOS mRNA W1DF

W1DF W2DF

29

iNOS eNOS

31

P4 StAR P450-scc 3β-HSD mRNA

W1DF W2DF P450-scc 3β-HSD mRNA W1DF

VEGF120 164 FGF-2 mRNA

VEGF164 mRNA W1DF

LH VEGF 45 P450-scc 106

W1DF LH LHr mRNA

W1DF

W1CL

4 W1DF W1CL

P4 VEGF

46

FGFR1

101

W1DF

W1CL

97

W1DF

W1CL

P4

32

58 P4 W1CL

P4

P4 P4

3 P4 mRNA

W1DF

W1DF W2DF

E2 LH

LHr

mRNA

W1DF

2 8

W1DF W2DF

LHr

mRNA

W1DF W1DF P4

Fig. 2.1.

Schematic diagram of experimental model of W1DF and W2DF in Experiment 1.

In the first follicular wave, the day of spontaneous ovulation was defined as Day

0. In the second follicular wave, the day of spontaneous ovulation was defined as

Day 7. Ovulation of the first-wave dominant follicle (DF) was induced by

GnRH treatment on Day 2 to induce a new follicular wave, and induced

ovulation was confirmed 36 h after GnRH treatment on Day 0. W1DF; DF in the

first follicular wave. W2DF;DF in the second follicular wave. B-mode US =

trans-rectal ultrasonography. Color Doppler US = trans-rectal color Doppler

ultrasonography.

W1DF

B-mode US Color Doppler US

Spontaneous ovulation B-mode US Color Doppler USInduced ovulation

W2DF

Day0 Day1 Day2 Day3 Day4 Day5 Day6


Spontaneous ovulation

Day 2Day 7

GnRH

33

Fig. 2.2.

Measuring method of blood flow area (BFA). Image of (A) is

dominant follicle with blood flow signals in the follicular wall

using color Doppler ultrasonography. The area of blood flow

signals which is enclosed by white line is measured as BFA

(image B).

(A) (B)

34

W1DF

W2DF

Fig. 2.3.

Comparative changes of plasma P4 concentration of W1DF and

W2DF from Day 0 to 6. Day 0 = The day of follicular wave

emergence. W1DF; Dominant follicle (DF) in the first follicular

wave. W2DF;DF in the second follicular wave. Effect of group

(G) (P < 0.001), day (D) (P < 0.001) and interaction between

group and day (G,D) (NS). Values are mean SEM of each point.

0

1.0

2.0

3.0

4.0

5.0

6.0

7.0

8.0


Pla

sma

P4

conc

entra

tion

of P

4(ng

/ml)

Days from follicular wave emergence

G P < 0.001D P < 0.001

G,D NS

Plas

ma

P 4 co

ncen

trat

ion

(ng/

ml)

35

Fig. 2.4.

Comparative changes of dominant follicle (DF) of W1DF and W2DF

from Day 3 to 6. Day 0 = The day of follicular wave emergence. W1DF;

DF in the first follicular wave. W2DF; DF in the second follicular wave.

Effect of group (G) (P < 0.01), day (D) (P < 0.01) and interaction

between group and day (G,D) (P < 0.01). The asterisk denotes

difference between each point, : P < 0.05. Values are mean SEM of

each point.

9

10

11

12

13

14

15

16

17

Day3 Day4 Day5 Day6

Dom

inan

t fol

licle

dia

met

er (m

m)


G P < 0.01

D P < 0.01

G,D P < 0.01

W1DFW2DF

8

36

Table 2.1. Blood flow in the follicular wall on Day6.

W1DF W2DF P-value

BFA(cm2) BF% (%)

0.32 0.05 0.21 0.05 24.37 2.64 18.35 3.17 P < 0.1

Values are mean SEM.

P < 0.1

W1DF = Dominant follicle (DF) in the first follicular wave. W2DF = DF in the second follicular wave. BFA = Blood flow area. BF% = The percentage of follicle circumference with blood flow signals.

37

Fig. 2.5.

Schematic diagram of experimental model of W1DF and W2DF in Experiment 2.

In the first follicular wave, the day of spontaneous ovulation was defined as Day

0. To induce follicular maturation, PGF2α and GnRH were administrated on Day

6 and 8, respectively. In the second follicular wave, the day of spontaneous

ovulation was defined as Day 7. Ovulation of the first-wave dominant follicle

(DF) was induced by GnRH treatment on Day 2 to induce a new follicular wave,

and induced ovulation was confirmed 36 h after GnRH treatment on Day 0.

PGF2α and GnRH were administrated on Day 6 and 8, respectively. W1DF; DF in

the first follicular wave. W2DF;DF in the second follicular wave. Color Doppler

US = trans-rectal color Doppler ultrasonography.

38

W1DF

Color Doppler US

Spontaneous ovulation Color Doppler US Induced ovulation

W2DF

Day0 Day9 Day6 Day7 Day8



Day 2 Day 7

PGF 2 GnRH

PGF 2 GnRH GnRH

Fig. 2.6.

Comparative changes of dominant follicle (DF) of W1DF and


emergence. W1DF; DF in the first follicular wave. W2DF;DF in

the second follicular wave. Effect of group (G) (P < 0.001), day

(D) (P < 0.001) and interaction between group and day (G,D) (NS).

Values are mean SEM of each point.

W1DF

W2DF


12

13

14

15

16

17

18

19

20

Day 6 Day 8 Day 9

G : P < 0.001 D : P < 0.001

G,D : NS Dom

inan

t fol

licle

dia

met

er (m

m)

39

W1DF

W2DF

0

1.0

2.0

3.0

4.0

5.0

6.0

7.0

8.0

Day6 Day7 Day8 Day9

Pla

sma

P4

conc

entra

tion

(ng/

ml)

G : P < 0.01D : P < 0.001

G,D : P < 0.05

P < 0.05


W1DF

W2DF

Fig. 2.7.

Comparative changes of plasma P4 concentration of W1DF and


emergence. W1DF; Dominant follicle (DF) in the first follicular

wave. W2DF;DF in the second follicular wave. Effect of group

(G) (P < 0.01), day (D) (P < 0.01) and interaction between

group and day (G,D) (P < 0.05). The asterisk denotes difference

between each point, : P < 0.05. Values are mean SEM of

each point.

G : P < 0.01 D : P < 0.01

G,D : P < 0.05

Pla

sma

P4 c

once

ntra

tion

(ng/

ml)

40

W1DF

W2DF

BF%

10

20

30

40

50

60

70

Day6 Day8 Day9 Days from follicular wave emergence

BF%

(%)

BFA

0.2

1.2

0.4

0.6

0.8

1.0

G : P < 0.01 D : P < 0.01

G,D : P < 0.05

Day6 Day8 Day9

G : P < 0.01 D : P < 0.01

G,D : P < 0.01

BFA

(cm

2 )

Fig. 2.8.

Comparative changes of blood flow area (BFA) and the percentage of follicle

circumference with blood flow signals (BF%) of W1DF and W2DF on Day 6 to 9.

Cows were treated with PGF2α and GnRH on Day 6 and Day 8, respectively. Day

0 = day of follicular wave emergence. W1DF; Dominant follicle (DF) in the first

follicular wave. W2DF;DF in the second follicular wave. For BFA, effect of

group (G) (P < 0.01), day (D) (P < 0.01) and interaction between group and day

(G,D)(P < 0.05). For BF%, effect of group (G) (P < 0.01), day (D) (P < 0.01) and

interaction between group and day (G,D) (P < 0.01). The asterisk denotes

difference between each points, : P < 0.05, : P < 0.01. Values are mean

SEM of each point.

41

Table 2.2. Plasma E2 concentration on Day 8.

W1DF W2DF P-value

E2 (pg/ml) 7.5 0.9 4.4 0.5


P < 0.01

Day8 = The day of after 48h of PGF2α treatment. W1DF = Dominant follicle (DF) in the first follicular wave. W2DF = DF in the second follicular wave.

42

Fig. 2.9.

Schematic diagram of experimental model of W1DF and W2DF in

Experiment 3. In the first follicular wave, the day of spontaneous ovulation

was defined as Day 0. To induce follicular maturation, PGF2α and GnRH

were administrated on Day 6 and 8, respectively. In the second follicular

wave, the day of spontaneous ovulation was defined as Day 7. Ovulation of

the first-wave dominant follicle (DF) was induced by GnRH treatment on

Day 2 to induce a new follicular wave, and induced ovulation was

confirmed 36 h after GnRH treatment on Day 0. PGF2α and GnRH were

administrated on Day 6 and 8, respectively. W1DF; DF in the first follicular

wave. W2DF;DF in the second follicular wave. DF were aspirated by trans-

vaginal ultrasonography in different estrous cycle.

43

W1DF

Spontaneous ovulation Induced ovulation

W2DF




Day 2 Day 7

PGF 2 GnRH

PGF 2 GnRH GnRH

Follicle aspilation

Follicle aspilation

Follicle aspilation

Follicle aspilation

Table 2.3. Primers used in real-time PCR.

Gene Sequence of nucleotide(5 - 3 ) Accesion No.

LHr Fa

Rb GCATCCACAAGCTTCCAGATGTTACGA GGGAAATCAGCGTTGTCCCATTGA

CYP19A1 F R

TTGCAAAGCATCCCCAGGTT AGGTCCACAACGGGCTGGTA

COX - 2 F R

TCCTGAAACCCACTCCCAACA TGGGCAGTCATCAGGCACAG

PGFS F R

GATCAAAGCGATTGCAGACA CAATGCGTTCAGGTGTCACT

PGES F R

CGCTGCTGGTCATCAAAAT GGAAGGGGTAGATGGTCTCC

iNOS F R

TCATCTTCGCCACCAAGCAG CAGTGATGGCCGACCTGATG

eNOS F R

GGAAATCGGGGGTCTGGAGT TTGGCGAGCTGAAAGCTGTG

Size(bp)

NM 174381

Z32741

AF031698

S54973

AY032727

U14640

M89952

204

151

241

113

186

180

219

Annealing temperature( C )

62

60

60

59

60

60

60

VEGF120 F R

CCCAGATGAGATTGAGTTCATTTT GCCTCGGCTTGTCACATTTT

M32976 377 56

VEGF164 F R

CCCAGATGAGATTGAGTTCATTTT AGCAAGGCCCACAGGGATTT

M32976 245 58

GAPDH F R

CTCTCAAGGGCATTCTAGGC TGACAAAGTGGTCGTTGAGG

NM001034034 160

a F, forward b R, reverse

60

StAR F R

GTGCATTTTGCCAATCACCT TTATTGAAAACGTGCCACCA

M174189 203 54.3

P450-scc F R

CTGCAAATGGTCCCACTTCT CACCTGGTTGGGTCAAACTT

K02103 209 56.3

3β-HSD F R

TCCACACCAGCACCATAGAA AAGGTGCCACCATTTTTCAG

X17614 118 56.3

44

Table 2.4. Sex steroids concentration in follicular fluid on Day 8.

W1DF W2DF P-value

P4 (ng/ml) E2 (ng/ml) A4 (ng/ml)

37.4 8.5 34.6 2.2 1,942.4 247.9 1,169.3 90.9

161.8 26.2 104.6 12.7

NS P < 0.05 P < 0.05

Values are mean SEM. Day 8 = The day of after 40h of PGF2α treatment. W1DF = Dominant follicle (DF) in the first follicular wave. W2DF = DF in the second follicular wave.

45

LHr CYP19A1

Fig. 2.10.

The relative amounts of mRNA of LHr and CYP19A1 in W1DF and

W2DF granulosa cells on Day 8. In each figure, the black bar indicates

W1DF and the white bar indicates W2DF. W1DF; Dominant follicle (DF) in

the first follicular wave. W2DF;DF in the second follicular wave. Day 8 =

the day of after 40h of PGF2α treatment. The asterisk denotes differences, *:

P < 0.05. Values are shown as mean SEM.

W1 DF

0.1

0

0.2

0.3

W 2DF 0

1.0

2.0

3.0

4.0

W1 DF W 2DF

/GAP

DH

/GAP

DH

*

46

W1DF W2DF P-value

P4 (ng/ml)

E2 (ng/ml)

90.1 11.8 91.4 7.7

82.5 20.7 64.1 8.7

NS

Table 2.5. Follicular fluid sex steroids concentration on Day 9.


NS

W1DF = Dominant follicle (DF) in the first follicular wave. W2DF = DF in the second follicular wave.

Day 9 = The day of after 24h of GnRH treatment.

47

/GA

PDH

W1 W20

0.04

0.08

0.12

0.16/G

APD

H

W20

0.2

0.4

0.6

0.8

W1

/GA

PDH

W1 W20

0.5

2.5

2.0

1.5

1.0

PGFS

COX-2 PGES

W1DF W2DF W1DF W2DF

W1DF W2DF Fig. 2.11.

The relative amounts of mRNA of COX-2, PGES and PGFS of W1DF

and W2DF granulosa cells on Day 9. In each figure, the black bar

indicates W1DF and the white bar indicates W2DF. W1DF; Dominant

follicle (DF) in the first follicular wave. W2DF;DF in the second

follicular wave. Day9 = the day of after 24h of GnRH treatment. Values

are shown as mean SEM.

48

iNOS eNOS

/GAP

DH

.0.012

0.08

0.04

0

W1 W2

/GA

PDH

0

0.005

0.01

0.015

0.02

0.025

W1 W2W1DF W2DF W1DF W2DF

Fig. 2.12.

The relative amounts of mRNA of iNOS and eNOS in W1DF and

W2DF granulosa cells on Day 9. In each figure, the black bar

indicates W1DF and the white bar indicates W2DF. W1DF;

Dominant follicle (DF) in the first follicular wave. W2DF;DF in the

second follicular wave. Day9 = the day of after 24h of GnRH

treatment. The asterisk denotes differences, : P < 0.1. Values are

shown as mean SEM.

49

0

0.5

1.0

1.5

2.0

2.5

/GA

PD

H

W1 W2

0

0.2

0.4

0.6

0.8

1.0

1.2

1.4

W1 W2

/GAP

DH

0

0.1

0.2

0.3

0.4

0.5

/GA

PD

HW1 W2

VEGF120 VEGF164

FGF-2

W1DF W2DF W1DF W2DF

W1DF W2DF

Fig. 2.13.

The relative amounts of mRNA of VEGF120, VEGF164 and FGF-2 in

W1DF and W2DF granulosa cells on Day 9. In each figure, the black

bar indicates W1DF and the white bar indicates W2DF. W1DF;

Dominant follicle (DF) in the first follicular wave. W2DF;DF in the

second follicular wave. Day9 = the day of after 24h of GnRH treatment.

The asterisk denotes differences, : P < 0.1, *: P < 0.05. Values are

shown as mean SEM.

*

50

0

5

10

15

20

25

30/G

AP

DH

W1 W2

02

468

101214

1618

/GA

PD

HW1 W2

0

20

40

60

80

100

120

140

/GA

PD

H

W1 W2

StAR P450-scc

3β-HSD

W1DF W2DF W1DF W2DF

W1DF W2DF

Fig. 2.14.

The relative amounts of mRNA of StAR, P450-scc, and 3 -HSD in W1DF

and W2DF granulosa cells on Day 9. In each figure, the black bar indicates

W1DF and the white bar indicates W2DF. W1DF; Dominant follicle (DF) in

the first follicular wave. W2DF;DF in the second follicular wave. Day9 = the

day of after 24h of GnRH treatment. The asterisk denotes differences, : P <

0.1, *: P < 0.05. Values are shown as mean SEM.

*

*

51

Fig. 2.15.

Schematic diagram of experimental model of W1CL and W2CL in Experiment 4.

Day 0 = The day of spontaneous ovulation. Day10 = The day of W1DF or W2DF

ovulation. W1DF; Dominant follicle (DF) in the first follicular wave. W2DF;DF

in the second follicular wave. W1CL; Corpus luteum (CL) formed after ovulation

of W1DF. W2CL; CL formed after ovulation of W2DF. B-mode US = trans-

rectal ultrasonography. Color Doppler US = trans-rectal color Doppler

ultrasonography.

Day10

Ovulation

Day11 Day13Day12 Day15Day14 Day19 Day20 Day21Day17Day16 Day18

Color Doppler US

W1CLW2CL

52

W1DF W2DF P-value

Diameter (mm) BF%(%)

Table 2.6. Diameter and blood flow in the follicular wall on Day 9.


P 0.01 17.7 0.3 15.7 0.5 55.3 4.0 36.1 2.3 P 0.01

W1DF = Dominant follicle (DF) in the first follicular wave. W2DF = DF in the second follicular wave.

Day 9 = The day of after 24h of GnRH treatment.

BF% = The percentage of follicle circumference with blood flow signals.

53

0

100

200

300

400

500

600

700

Cro

ss-s

ectio

nal a

rea

of c

orpu

s lu

teum

(mm

2 )

12 15 18 21

G : P < 0.01 D : P < 0.01

G,D : P < 0.05

W1CL W2CL


Fig. 2.16.

Comparative changes of cross-sectional area of corpus luteum (CL) of

W1CL and W2CL on Day 12, 15, 18 and 21. Day 10 = ovulation day of

W1DF and W2DF. W1DF; Dominant follicle (DF) in the first follicular

wave. W2DF;DF in the second follicular wave. W1CL; CL formed after

ovulation of W1DF. W2CL; CL formed after ovulation of W2DF. Effect

of group (G) (P < 0.01), day (D) (P < 0.01) and interaction between

group and day (G,D)(P < 0.05). The asterisk denotes difference between

each point, : P < 0.05. Values are mean SEM of each point.

54

0

100

200

300

400

500

12 15 18 21 Days from follicular wave emergence

Blo

od fl

ow a

rea

of c

orpu

s lu

teum

(mm

2 )

G : P < 0.05 D : P < 0.05

G,D : P < 0.05

W1CL W2CL

Fig. 2.17. Comparative changes of blood flow area in the corpus luteum (CL) of

W1CL and W2CL on Day 12, 15, 18 and 21. Day 10 = ovulation

day of W1DF and W2DF. W1DF; Dominant follicle (DF) in the first

follicular wave. W2DF;DF in the second follicular wave. W1CL; CL

formed after ovulation of W1DF. W2CL; CL formed after ovulation

of W2DF. Effect of group (G) (P < 0.05), day (D) (P < 0.05) and

interaction between group and day (G,D)(P < 0.05). The asterisk

denotes difference between each point, : P < 0.05. Values are mean

SEM of each point.

55

0

1.0

2.0

3.0

4.0

5.0

6.0

7.0

10 14 16 17 18 20 21 13 12 11 15 19

Pla

sma

P4 c

once

ntra

tion

(ng/

ml)

G:P<0.05 D:P<0.05

GD:NS


W1CL W2CL

G : P < 0.05 D : P < 0.05

G,D : NS

Fig. 2.18.

Comparative changes in the plasma P4 concentration of W1CL and

W2CL. Blood samples were collected daily from Day 10 to 21. Day

10 = ovulation day of W1DF and W2DF. W1DF; Dominant follicle

(DF) in the first follicular wave. W2DF;DF in the second follicular

wave. W1CL; Corpus luteum (CL) formed after ovulation of W1DF.

W2CL; CL formed after ovulation of W2DF. Effect of group (G) (P <

0.05), day (D) (P < 0.05) and interaction between group and day

(G,D)(NS). Values are mean SEM of each point.

56

57

1 W1DF

58

3 1

2 W1DF

W2DF

P4 LH 15

52 Adams 5 P4 W1DF

W1DF

P4

W1DF

P4

W1DF

W1DF

40

33 W1DF W1DF

W1DF

3 2 1 7 W1DF P4

W1DF W1DF

3 2 1

59

30

6.1 0.4 ± ) 44

TMR

3 2 2

12

Day 0 Day 6 W1DF

W1DF

29

3 2 3

5.0MHz

HS 101V

3 2 4

21G TERUMO

10ml 4

2,000 g 20 P4 30

60

3 2 5 P4

Day 6 P4 EIA 2

5.2 7.4%

3 2 6

Day 6 P4 2

Pearson

± Day 6

P4 F Student

t P < 0.05

StatView Abacus Concepts Inc. Berkeley CA USA

3 3 2 W1DF 5

W1DF P4

3 3 1

5

5.6 1.0 ± )

TMR

61

3 3 2

12

Day 0 Day 0 4

W1DF Day 4 W1DF

W1DF

Day 0 4 Day

4 W1DF

5 n

5 n 5 5 1

3 3 3

5.0MHz

HS 101V

2 2×3.14

3 3 4

21G TERUMO

10ml 4

62

2,000 g 20 P4 E2 30

3 3 5

2

3 3 6 P4 E2 A4

2 P4

E2 A4 85 81 84 P4

5.5% 7.4% E2 6.2

8.6% A4 5.6 7.8%

3 3 7 P4 E2

Day 0 4 P4 Day 4 E2 EIA

2 P4 5.2

7.4% E2 6.0 8.4

3 3 8

± P4

2

Bartlet

Post-hoc Schefe Day 4

E2 P4 E2 A4 F

63

Student t P < 0.05


3 4

3 4 1 1

n 22 6.2±0.6 n 22 6.2±0.4

n 29 6.1±0.4 P4 3.0±0.2

ng/ml n 16 6.0±0.7 P4

n 13 6.2±0.6 P4

Table 3.1.

Day 6 P4

Fig. 3.1.

P4

Fig. 3.2.

Day 6

Table 3.2.

3 4 2 2

Fig. 3.3. Day 2

4

P4 Fig. 3.4. Fig. 3.5.

P4

64

Day 4 E2 Day 4

Table 3.4. E2

A4 P4

3 5

1 Day 6 P4 W1DF

W1DF

P4 2

E2 P4 W1DF

2 W1DF W2DF P4

Day 6 W1DF P4

P4

W1DF

5

P4

P4 W1DF

P4 2 W2DF

P4

Day 6 P4 W1DF

Ginther 42 W1DF

W1DF

65

W1DF W1DF

W1DF

42

2 42 84 86 W1DF

LH 109

Day 6 P4

W1DF

2 W1DF

W1DF P4

2 Day 0 4

P4 P4

1 W1DF

5 1 7

Day 4 W1DF

P4

E2 A4

66

P4 22

P4 A4 31 59 A4

E2 12

A4 E2

P4

P4

75

P4

E2

Ginther 42 W1DF

W1DF

W1DF

E2

3 6

W1DF

W1DF 1

67

7 W1DF P4

W1DF W1DF

P4

W1DF P4

2 5

P4

P4

P4

P4

Table 3.1. Plasma P4 concentration on Day 6.

IG CG P-value

P4 (ng/ml) 2.7 0.3 3.5 0.3 P < 0.05

68


Day0 = The day of the first follicular wave emergence.

IG; Ipsilateral group: Dominant follicle (DF) ipsilateral to the corpus luteum (CL). CG; Contralateral group: DF contralateral to the CL.

Fig. 3.1.

The linear relationship between plasma P4 concentration and

diameter of the first wave dominant follicle on Day 6. Day 0 = The

day of the first follicular wave emergence.

R=0.23 P:NS

1 .0

2 .0

3 .0

4 .0

5 .0

6 .0

7 .0

13 14 15 16 17 18

Pla

sma

P4 c

once

ntra

tion

(ng/

ml)

Diameter of the first wave dominant follicle (mm)

69

Fig. 3.2.

The linear relationship between plasma P4 concentration and

diameter of the first wave dominant follicle (DF) on Day6 in IG and

CG. Day0 = The day of the first follicular wave emergence. IG;

Ipsilateral group: DF ipsilateral to the corpus luteum (CL). CG;

Contralateral group: DF contralateral to the CL.

R=0.36 P:NS

R=0.12

P:NS

1 .0

2 .0

3 .0

4 .0

5 .0

6 .0

7 .0

13 14 15 16 17 18

IG C G

Pla

sma

P4 c

once

ntra

tion

(ng/

ml)

Diameter of the first wave dominant follicle (mm)

70

Table 3.2. Diameter of the first follicular wave dominant follicle on Day6.

IG CG P-value

Diameter (mm) 15.5 0.2 14.7 0.2 P < 0.01

71




Fig. 3.3.

Comparative changes of the first follicular wave dominant follicle (DF) of

IG and CG from Day 0 to 4. Day 0 = The day of the first follicular wave

emergence. Effect of group (G) (NS), day (D) (P < 0.01) and interaction

between group and day (G,D) (NS). IG; Ipsilateral group: DF ipsilateral to

the corpus luteum (CL). CG; Contralateral group: DF contralateral to the CL.


0

2

4

6

8

10

12

14


IG C G

G : NS D : P < 0.001

G,D : NS

Dia

met

er o

f dom

inan

t fol

licle

(mm

)


72

0

50

100

150

200

250

300


Cro

ss-s

ectio

nal a

rea

of c

orpu

s lu

teum

(mm

2 )


IGCG

G : NSD : P < 0.001

G,D : NS

Fig. 3.4.

Comparative changes of cross-sectional area of corpus luteum (CL) of IG

and CG from Day 0 to 4. Day 0 = The day of the first follicular wave

emergence. Effect of group (G) (NS), day (D) (P < 0.001) and interaction

between group and day (G,D) (NS). IG; Ipsilateral group: Dominant follicle

(DF) ipsilateral to the CL. CG; Contralateral group: DF contralateral to the

CL. Values are mean SEM of each point.


73

Fig. 3.5.

Comparative changes of plasma P4 concentration of IG and CG from Day

0 to 4. Day 0 = The day of the first follicular wave emergence. Effect of

group (G) (NS), day (D) (P<0.001) and interaction between group and day

(G,D) (NS). IG; Ipsilateral group: Dominant follicle (DF) ipsilateral to the

corpus luteum (CL). CG; Contralateral group: DF contralateral to the CL.


0

0.5

1 .0

1.5

2.0

2.5

3 .0


IG C G

G : NS D : P < 0.001

G,D : NS

Pla

sma

P4 c

once

ntra

tion

(ng/

ml)


74

Table 3.3. Plasma E2 concentration on Day4.

IG CG P-value

E2 (pg/ml) 1.69 0.12 1.51 0.15 P < 0.1

75




Table 3.4. Sex steroids concentration in follicular fluid on Day4.

IG CG P-value

P4 (ng/ml) E2 (ng/ml) A4 (ng/ml)

48.8 12.7 16.2 3.2 419.2 33.2 339.4 53.5

84.6 23.1 102.7 40.2

P < 0.05 NS NS



IG; Ipsilateral group: Dominant follicle (DF) ipsilateral to the corpus luteum (CL).

76

CG; Contralateral group: DF contralateral to the CL.

77

1 W1DF

78

4 1

3 W1DF

W1DF W1DF

P4

20

E2

99

W1DF

4 2 1 W1DF W1DF

P4

4 2 1

n 30 2.2 0.2

99.9 8.0 37.6 1.6 kg

4 2 2

24

Day 0 Day 2 4 6 8 10

79

2

4 2 3

W1DF 5.0MHz

HS 101V

W1DF W1DF

4 2 4

5.0MHz

HS 101V

2 2 3.14

4 2 5

21G TERUMO

10ml 4 2,000 g

20 P4 30

80

4 2 6 P4

Day 2 4 6 8 10 P4 EIA

2 P4 5.2

7.4%

4 2 7

P4

2

Bartlet Pos-hoc

Sheffe P < 0.05


4 3 2 W1DF

4 3 1

153 75

2008 3 2013 9

238

119.8 5.2 2.2 0.1 33.9 0.5 kg/

81

BCS 2.91 0.02 637.4 4.6 kg;

112 14.2 0.3 440.6 5.0 kg)

TMR

1 2

60 350

4 3 2

24

24 2

Day 0

Day 4 8 W1DF

2

40

4 3 3

W1DF 5.0MHz

HS 101V

W1DF W1DF

82

4 3 4

4 3 5

W1DF 6 9 10 5

441kg 441kg

120 120

34kg/ 34kg/ 1 2 3 BCS 3

3 27 637kg 637kg

1 BCS

BCS

2 P < 0.05

R version 2.14.0 for Mac OS X.

4 4

83

4 4 1 1

n 16 104.8 10.5 1.9 0.2

35.5 2.1 kg/ n 14 94.4 12.4 2.6

0.4 39.9 2.3 kg/ Fig. 4.1.

Fig. 4.2. Day 8

Day 4 6 8

Day 4 8

P4

Fig. 4.3

4 4 2 2

54.0 189/350 48.9

122/238 58.9 67/112

n 184 n 166

n 129 115.4 5.7 2.4

0.1 34.9 0.7 kg/ 646.7 6.3 kg BCS 2.91 0.03

n 109 118.3 5.6 2.1 0.1 32.7

0.7 kg/ 626.6 6.5 kg BCS 2.92 0.03

n 55 434.1 6.3 kg 14.1 0.6 n

57 446.9 8.0 kg 14.2 0.5

vs. 40.2 74/184 vs. 69.3

115/166 Fig. 4.4.

84

vs. 38.0 49/129 vs. 67.0 73/109 vs. 45.5

25/55 vs. 73.7 42/57

BCS

4 5

1 W1DF

W1DF

P4

P4 W1DF

3 Day 6 W1DF P4

P4

P4 W1DF

3 2

P4 78

P4

23

100

23 97

P4

85

3 1

P4

W1DF W1DF

40 42

3 2 Day 4

1

Day 4 3 2 Day 6

3 1 Day 8

W1DF

P4 W1DF

W1DF P4

W1DF

42

2 42

84 86

W1DF W1DF

109

W1DF

W1DF

86

W1DF

3 1 2 1 W1DF

P4 20

E2

99

91

P4

A4 E2

47 2

W1DF

E2 1 E2

W1DF E2 E2

79 W1DF

E2 in vitro

E2 P4 mRNA

96 in vitro E2

P4 102 E2

W1DF

W1DF E2

87

W1DF

P4

2

P4 W1DF

3

1 P4

W1DF P4 W1DF

W1DF

W1DF

4 6

W1DF

W1DF P4

W1DF

W1DF

100

200

300

400

500

600

700


IGCG

G : NSD : P < 0.001

G,D : NS


Cro

ss-s

ectio

nal a

rea

of c

orpu

s lu

teum

(mm

2 )

Fig. 4.1.

Comparative changes of corpus luteum (CL) of IG and CG on Day 2, 4, 6, 8,

10. Day 0 = The day of the first follicular wave emergence. Effect of group (G)

(NS), day (D) (P < 0.001) and interaction between group and day (G,D) (NS).

IG; Ipsilateral group: Dominant follicle (DF) ipsilateral to the CL. CG;

Contralateral group: DF contralateral to the CL. Values are mean SEM of

each point.

88

6

8

10

12

14

16

18

20


IG CG

G : P < 0.01 D : P < 0.001

G,D : P < 0.01


Dia

met

er o

f dom

inan

t fol

licle

(mm

)

Fig. 4.2.

Comparative changes of the first follicular wave dominant follicle (DF)

of IG and CG on Day 2, 4, 6, 8, 10. Day 0 = The day of the first

follicular wave emergence. Effect of group (G) (P < 0.01), day (D) (P <

0.001) and interaction between group and day (G,D) (P < 0.01). IG;

ipsilateral group: DF ipsilateral to the corpus luteum (CL). CG;

contralateral group: DF contralateral to the CL. The asterisk denotes

difference between each point, : P < 0.05. Values are mean SEM of

each point.

89

0

1.0

2.0

3.0

4.0

5.0

6.0

7.0


IGCG

G : NSD : P < 0.001

G,D : NS


Pla

sma

conc

entra

tion

of P

4(n

g/m

l)

Fig. 4.3.

Comparative changes of plasma P4 concentration of IG and CG on

Day 2, 4, 6, 8, 10. Day 0 = The day of the first follicular wave

emergence. Effect of group (G) (NS), day (D) (P < 0.001) and

interaction between group and day (G,D) (NS). IG; ipsilateral group:

Dominant follicle (DF) ipsilateral to the corpus luteum (CL). CG;

contralateral group: DF contralateral to the CL. Values are mean

SEM of each point.

Pla

sma

P4 c

once

ntra

tion

(ng/

ml)


90

0

10

20

30

40

50

60

70

80

Con

cept

ion

rate

(%)

Lactating dairy cows Dairy heifers

38.0% (49/129)

67.0% (73/109)

45.5% (25/55)

73.7% (42/57)

IG CG

Figure 4.4. Conception rates in IG and CG. Conception rates were

different between IG and CG in both lactating dairy cows

and dairy heifers. IG; ipsilateral group: Dominant follicle

(DF) ipsilateral to the corpus luteum (CL). CG;

contralateral group: DF contralateral to the CL. The asterisk

denotes difference between each point, : P < 0.01.

91

92

93

1 1

2 3

1 W1DF

W1DF

W1DF

E2 P4 E2 A4

P4

2 W1DF 2

W2DF W1DF n 7 W2DF n 7 1

Day 3 6 Day 0 P4

Day 6

W1DF W2DF Day 6

W1DF

P4

94

W1DF P4 W2DF P4

W1DF

P4

P4

LHr

LH P4

LH

W1DF LH

W1DF

W1DF W2DF

2 PGF2α Day 6 GnRH Day 8

W1DF W2DF GnRH

Day 8 E2 W1DF

W1DF GnRH

W1DF GnRH W1DF

1

GnRH E2

GnRH 26

W1DF

3 W1DF GnRH

W1DF n 7 W2DF n 7 24 W1DF n 6 W2DF n 6

95

mRNA

GnRH 2 E2

E2 A4 CYP19A1 mRNA

W1DF GnRH 24 LH

LHr mRNA

E2 A4 W1DF

CYP19A1 mRNA A4

E2

E2 A4

W1DF GnRH E2

LHr mRNA W1DF LH

LHr mRNA W1DF P4

LH

LHr mRNA

GnRH 24 mRNA LH

E2

P4 VEGF

FGF

LH PG PGF2α

PGE2 PGE2

NO iNOS eNOS

LH

W1DF

96

COX-2 PGES PGFS mRNA

LH

W1DF W2DF

iNOS mRNA eNOS mRNA W1DF

W1DF W2DF

P4 P450-scc 3β-HSD mRNA

VEGF164 mRNA W1DF

P4

W1DF LH

4 W1DF W2DF W1DF

n 6 W2DF n 6 P4 W1DF

P4

W1DF W2DF

W1DF P4

W1DF

P4 P4

P4

W1DF W2DF

97

3 W1DF

1 44 Day 6

P4

2 P4

Day 6 P4

Day 6 P4 W1DF

P4 2 5

n 5 n 5

Day 0 4 P4 Day 4

E2 E2 A4 P4

P4 P4 1

1 Day 6 P4

E2

E2 A4

P4 W1DF

W1DF

E2

E2 P4 P4

98

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1. Acosta, T.J., Berisha, B., Ozawa, T., Sato, K., Schams, D. and Miyamoto, A. (1999).

Evidence for a local endothelin-angiotensin-atrial natriuretic peptide systemin bovine

mature follicles in vitro: effects on steroid hormones and prostaglandin secretion. Biol.

Reprod. 61, 1419~1425.

2. Acosta, T.J., Hayashi, K.G., Matsui, M. and Miyamoto, A. (2005). Changes in

follicular vascularity during the first follicular wave in lactating cows. J. Reprod. Dev.

51, 273~280.

3. Acosta, T.J., Miyamoto, A., Ozawa, T., Wijayagunawardane, M.P. and Sato, K. (1998).

Local release of steroid hormones, prostaglandin E2, and endothelin-1 from bovine

mature follicles In vitro: effects of luteinizing hormone, endothelin-1, and cytokines.

Biol. Reprod. 59, 437~443.

4. Adams, G.P., Jaiswal, R., Singh, J. and Malhi, P. (2008). Progress in understanding

ovarian follicular dynamics in cattle. Theriogenology. 69, 72~80.

5. Adams, G.P., Matteri, R.L. and Ginther, O.J. (1992). Effect of progesterone on ovarian

follicles, emergence of follicular waves and circulating follicle-stimulating hormone in

heifers. J. Reprod. Fertil. 96, 627~640.

6. Adams, G.P., Matteri R.L., Kastelic, J.P., Ko, J.C. and Ginther, O.J. (1992). Association

between surges of follicle-stimulating hormone and the emergence of follicular waves

in heifers. J. Reprod. Fertil. 94, 177~188.

7. Ahmad, N., Townsend, E.C., Dailey, R.A. and Inskeep, E.K. (1997). Relationships of

hormonal patterns and fertility to occurrence of two or three waves of ovarian follicles,

102

before and after breeding, in beef cows and heifers. Anim. Reprod. Sci. 49, 13~28.

8. Alila, H.W. and Hansel, W. (1984). Origin of different cell types in the bovine corpus

luteum as characterized by specific monoclonal antibodies. Biol. Reprod. 31,

1015~1025.

9. Badinga, L., Driancourt, M.A., Savio, J.D., Wolfenson, D., Drost, M., De La Sota, R.L.

and Thatcher, W.W. (1992). Endocrine and ovarian responses associated with the

first-wave dominant follicle in cattle. Biol. Reprod. 47, 871~883.

10. Badinga, L., Thatcher, W.W., Diaz, T., Drost, M. and Wolfenson, D. (1993). Effect of

environmental heat stress on follicular development in lactating cows. Theriogenology.

39, 797~810.

11. Baird, D.T., Land, R.B., Scaramuzzi, R.J. and Wheeler, A.G. (1976). Endocrine

changes associated with luteal regression in the ewe; the secretion of ovarian oestradiol,

progesterone and androstenedione and uterine prostaglandin F2 throughout the

oestrous cycle. J. Endocrinol. 69, 275~286.

12. Bao, B. and Garverick, H.A. (1998). Expression of steroidogenic enzyme and

gonadotropin receptor genes in bovine follicles during ovarian follicular waves: a

review. J. Anim. Sci. 76, 1903~1921.

13. Beg, M.A., Bergfelt, D.R., Kot, K., Wiltbank, M.C. and Ginther, O.J. (2001).

Follicular-fluid factors and granulosa-cell gene expression associated with follicle

deviation in cattle. Biol. Reprod. 64, 432~441.

14. Bello, N.M., Steibel, J.P. and Pursley, J.R. (2006). Optimizing ovulation to first GnRH

improved outcomes to each hormonal injection of ovsynch in lactating dairy cows. J.

Dairy Sci. 89, 3413~3424.

15. Bergfeld, E.G., Kojima, F.N., Cupp, A.S., Wehrman, M.E., Peters, K.E., Mariscal, V.,

103

Sanchez, T. and Kinder, J.E. (1996). Changing dose of progesterone results in sudden

changes in frequency of luteinizing hormone pulses and secretion of 17β-estradiol in

bovine females. Biol. Reprod. 54, 546~553.

16. Bleach, E.C.L., Glencross, R.G. and Knight, P.G. (2004). Association between ovarian

follicle development and pregnancy rates in dairy cows undergoing spontaneous

oestrous cycles. Reproduction. 127, 621~629.

17. Burns, D.S., Krassel, F.J.,Ireland, J.L.H., Knight, P.G. and Ireland, J.J. (2005). Number

of antral follicles during follicular waves in cattle: evidence for high variation among

animals, very high repeatability in individuals, and an inverse association with serum

follicle-stimulating hormone concentrations. Biol. Reprod. 73, 54~62.

18. Butler, W.R. (1998). Review: Effect of protein nutrition on ovarian and uterine

physiology in dairy cattle. J Dairy Sci. 81, 2533~2539.

19. Celik, H.A., Aydm, I., Sendag, S. and Dinc, D.A. (2005). Number of follicular waves

and their effect on pregnancy rate in the cow. Reprod. Domest. Anim. 40, 87~92.

20. Cerbito, W.A., Miyamoto, A., Balagapo, C.R. Jr., Natural, N.G., Miyazawa, K. and

Sato, K. (1994). Prostaglandin E2 levels in uterine tissues and its relationship with

uterine and luteal progesterone during the estrous cycle in dairy cows. Theriogenology.

42, 941~950.

21. Daray, F.M., Minvielle, A.I., Puppo, S. and Rothlin, R.P. (2004). Vasoconstrictor effects

of 8-iso-prostaglandin E2 and 8-iso-prostaglandin F2 on human umbilical vein. Eur. J.

Pharmacol. 499, 189~195.

22. Diaz, F.J., Anderson, L.E., Wu, Y.L., Rabot, A., Tsai, S.J. and Wiltbank, M.C. (2002).

Regulation of progesterone and prostaglandin F2 production in the CL. Mol. Cell.

Endocrinol. 31, 65~80.

104

23. Endo, N., Nagai, K., Tanaka, T. and Kamomae, H. (2012). Comparison between

lactating and non-lactating dairy cows on follicular growth and corpus luteum

development, and endocrine patterns of ovarian steroids and luteinizing hormone in the

estrous cycles. Anim. Reprod. Sci. 134, 112~118.

24. Endo, N., Nagai, K., Tanaka, T. and Kamomae, H. (2012). Profiles of ovarian steroids,

luteinizing hormone and estrous signs from luteolysis to ovulation in lactating and

non-lactating dairy cows. J. Reprod. Dev. 58, 685~690.

25. Espey LL. (1980). Ovulation as an inflammatory reaction-a hypothesis. Biol. Reprod.

22, 73~106.

26. Espey, L.L. (1994). Current status of the hypothesis that mammalian ovulation is

comparable to an inflammatory reaction. Biol. Reprod. 50, 233~238.

27. Ferguson, J.D., Galligan, D.T. and Thomsen, N. (1994). Principal descriptors of body

condition score in Holstein cows. J. Dairy Sci. 77, 2695~2703.

28. Figueiredo, R.A., Barros, C.M., Pinheiro, I.O.L. and Sole, J.M.P. (1997). Ovarian

follicular dynamics in nelore breed (Bos indicus) cattle. Theriogenology. 47,

1489~1505.

29. Findlay, J.K. (1986). Angiogenesis in reproductive tissues. J. Endocrinol. 111,

357~366.

30. Forde, N., Beltman, M.E., Lonergan, P., Diskin, M., Roche, J.F. and Crowe, M.A.

(2011). Oestrous cycles in Bos taurus cattle. Anim. Reprod. Sci. 124, 163~169.

31. Fortune, J.E. (1986). Bovine theca and granulosa cells interact to promote androgen

production. Biol. Reprod. 35, 292~299.

32. Garcia-Ispierto, I., Roselló, M.A., De Rensis, F. and López-Gatius, F. (2013). A

five-day progesterone plus eCG-based fixed-time AI protocol improves fertility over

105

spontaneous estrus in high-producing dairy cows under heat stress. J. Reprod. Dev. 17,

544~548.

33. Ginther, O.J. and Hoffman, M.M. (2014). Intraovarian effect of dominant follicle and

corpus luteum on number of follicles during a follicular wave in heifers.

Theriogenology. 82, 169~175.

34. Ginther, O.J., Beg, M.A., Bergfelt, D.R., Donadeu, F.X. and Kot, K. (2001). Follicle

selection in monovular species. Biol. Reprod. 65, 638~647.

35. Ginther, O.J., Beg, M.A., Donadeu, F.X. and Bergfelt, D.R. (2003). Mechanism of

follicle deviation in monovular farm species. Anim. Reprod. Sci. 78, 239~257.

36. Ginther, O.J., Bergfelt, D.R., Beg, M.A. and Kot, K. (2001). Follicle selection in cattle:

role of luteinizing hormone. Biol. Reprod. 64, 197~205.

37. Ginther, O.J., Bergfelt, D.R., Beg, M.A. and Kot, K. (2001). Follicle selection in cattle:

relationships among growth rate, diameter ranking, and capacity for dominance. Biol.

Reprod. 65, 345~350.

38. Ginther, O.J., Bergfelt, D.R., Kulick, L.J. and Kot, K. (2000). Selection of the dominant

follicle in cattle: role of estradiol. Biol. Reprod. 63, 383~389.

39. Ginther, O.J., Kastelic, J.P. and Knopf, L. (1989). Composition and characteristics of

follicular waves during the bovine estrous cycle. Anim. Reprod. Sci. 20, 187~200.

40. Ginther, O.J., Kastelic, J.P. and Knopf, L. (1989). Intraovarian relationships among

dominant and subordinate follicles and the corpus luteum in heifers. Theriogenology.

32, 787~795.

41. Ginther, O.J., Knopf, L. and Kastelic, J.P. (1989). Temporal associations among ovarian

events in cattle during oestrous cycles with two and three follicular waves. J. Reprod.

Fertil. 87, 223~230.

106

42. Ginther, O.J., Rakesh, H.B. and Hoffman, M.M. (2014). Blood flow to follicles and CL

during development of the periovulatory follicular wave in heifers. Theriogenology. 15,

304~311.

43. Ginther, O.J., Wiltbank, M.C., Fricke, P.M., Gibbons, J.R. and Kot, K. (1996).

Selection of the dominant follicle in cattle. Biol Reprod. 55, 1187~1194.

44. (2010). 3) .

, pp.

291~293. ,

45. Hazzard, T.M., Molskness, T.A., Chaffin, C.L. and Stouffer, R.L. (1999). Vascular

endothelial growth factor (VEGF) and angiopoietin regulation by gonadotrophin and

steroids in macaque granulosa cells during the peri-ovulatory interval. Mol. Hum.

Reprod. 5, 1115~1121.

46. Hazzard, T.M., Xu, F. and Stouffer, R.L. (2002). Injection of soluble vascular

endothelial growth factor receptor 1 into the preovulatory follicle disrupts ovulation

and subsequent luteal function in rhesus monkeys. Biol. Reprod. 67, 1305~1312.

47. Hunter, R.H., Cook, B. and Poyser, N.L. (1983). Regulation of oviduct function in pigs

by local transfer of ovarian steroids and prostaglandins: a mechanism to influence

sperm transport. Eur. J. Obstet. Gynecol. Reprod. Biol. 14, 225~232.

48. Jaiswal, R.S., Singh, J., Marshall, L. and Adams, G.P. (2009). Repeatability of 2-wave

and 3-wave patterns of ovarian follicular development during the bovine estrous cycle.


49. (2012). . , ,

4 pp. 49~55. ,

50. Kawate, N., Itami, T., Choushi, T., Saitoh, T., Wada, T., Matsuoka, K., Uenaka, K.,

107

Tanaka, N., Yamanaka, A., Sakase, M., Tamada, H., Inaba, T. and Sawada, T. (2004).

Improved conception in timed-artificial insemination using a progesterone-releasing

intravaginal device and Ovsynch protocol in postpartum suckled Japanese Black beef

cows. Theriogenology. 15, 399~406.

51. Kindahl, H., Edqvist, L.E., Granstrom, E. and Bane, A. (1976). The release of

prostaglandin F2 as reflected by 15-keto-13,14-dihydroprostaglandin F2 in the

peripheral circulation duringnormal luteolysis in heifers. Prostaglandins. 11, 871~878.

52. Kojima, N., Stumpf, T.T., Cupp, A.S., Werth, L.A., Roberson, M.S., Wolfe, M.W.,

Kittok, R.J. and Kinder, J.E. (1992). Exogenous progesterone and progestins as used in

estrous synchrony regimens do not mimic the corpus luteum in regulation of luteinizing

hormone and 17β-estradiol in circulation of cows. Biol. Reprod. 47, 1009~1017.

53. Kulick, L.J., Kot, K., Wiltbank, M.C. and Ginther, O.J. (1999). Follicular and hormonal

dynamics during the first follicular wave in heifers. Theriogenology. 52, 913~921.

54. Lamothe, P., Bousquet, D. and Guay, P. (1977). Cyclic variation of F prostaglandins in

the uterine fluids of the cow. J. Reprod. Fertil. 50, 381~382.

55. Lopez, H., Satter, L.D. and Wiltbank, M.C. (2004). Relationship between level of milk

production and estrous behavior of lactating dairy cows. Anim. Reprod. Sci. 81,

209~223.

56. Luo, W., Gumen, A., Haughian, J.M. and Wiltbank, M.C. (2011). The role of

luteinizing hormone in regulating gene expression during selection of a dominant

follicle in cattle. Biol. Reprod. 84, 369~378.

57. Lüttgenau, J. and Bollwein, H. (2014). Evaluation of bovine luteal blood flow by using

color Doppler ultrasonography. Reprod. Biol. 14, 103~109.

58. Lüttgenau, J., Ulbrich, S.E., Beindorff, N., Honnens, A., Herzog, K. and Bollwein, H.

108

(2011). Plasma progesterone concentrations in the mid-luteal phase are dependent on

luteal size, but independent of luteal blood flow and gene expression in lactating dairy

cows. Anim. Reprod. Sci. 125, 20~29.

59. Magoffin, D.A. (2005). Ovarian theca cell. Int. J. Biochem. Cell. Biol. 37, 1344~1349.

60. Martínez, M.F., Kastelic, J.P., Adams, G.P., Janzen, E., McCartney, D.H. and Mapletoft,

R.J. (2000). Estrus synchronization and pregnancy rates in beef cattle given CIDR-B,

prostaglandin and estradiol, or GnRH. Can. Vet. J. 41, 786~790.

61. McClellan, M.C., Dieckman, M.A., Abel, J.H. Jr. and Niswender, G.D. (1975).

Luteinizing hormone, progesterone and the morphological development of normal and

superovulated corpora lutea in sheep. Cell Tissue Res. 164, 291~307.

62. Mihm, M. and Bleach, E.C. (2003) Endocrine regulation of ovarian antral follicle

development in cattle. Anim. Reprod. Sci. 78, 217~237.

63. (2011). . pp.

370~378. ,

64. Miyamoto, A., Okuda, K., Schweigert, F.J. and Schams, D. (1992). Effects of basic

fibroblast growth factor, transforming growth factor-β and nerve growth factor on the

secretory function of the bovine corpus luteum in vitro. J. Endocrinol. 135, 103~114.

65. Moenter, S.M., Brand, R.C. and Karsch, F.J. (1992). Dynamics of gonadotropin

releasing hormone (GnRH) secretion during the GnRH surge: insights into the

mechanism of GnRH surge induction. Endocrinology. 130, 2978~2984.

66. Murphy, M.G., Enright, W.J., Crowe, M.A., McConnell, K., Spicer, L.J., Boland, M.P.

and Roche, J.F. (1991). Effect of dietary intake on pattern of growth of dominant

follicles during the oestrous cycle of beef heifers. J. Reprod. Fertil. 92, 333~338.

67. Nett, T.M., Staigmiller, R.B., Akbar, A.M., Diekman, M.A., Ellinwood, W.E. and

109

Niswender, G.D. (1976). Secretion of prostaglandin F2 in cycling and pregnant ewes. J.

Anim. Sci. 42, 876~880.

68. Niswender, G.D., Juengel, J.L., McGuire, W.J, Belfiore, C.J. and Wiltbank, M.C.

(1994). Luteal function: the estrous cycle and early pregnancy. Biol. Reprod. 50,

239~247.

69. Noseir W.M.B. (2003). Ovarian follicular activity and hormonal profile during estrous

cycle in cows: the development of 2 versus 3 waves. Reprod. Biol. Endocrinol.1:50.

70. Perry, G.A., Smith, M.F., Lucy, M.C., Green, J.A., Parks, T.E., MacNeil, M.D., Roberts,

A.J. and Geary, T.W. (2005). Relationship between follicle size at insemination and

pregnancy success. Proc. Natl. Acad. Sci. 102, 5268~5273.

71. Peters, M.W., Pursley, J.R. and Smith, G.W. (2004). Inhibition of intrafollicular PGE2

synthesis and ovulation following ultrasound-mediated intrafollicular injection of the

selective cyclooxygenase-2 inhibitor NS-398 in cattle. J. Anim. Sci. 2004 82,

1656~1662.

72. Pursley, J.R., Mee, M.O. and Wiltbank, M.C. (1995). Synchronization of ovulation in

dairy cows using PGF2 and GnRH. Theriogenology. 44, 915~923.

73. Rahe, C.H., Owens, R.E., Fleeger, J.L., Newton, H.J. and Harms, P.G. (1980). Pattern

of plasma luteinizing hormone in the cyclic cow: dependence upon the period of the

cycle. Endocrinology. 107, 498~503.

74. Rajamahendran, R. and Taylor, C. (1990). Characterization of ovarian activity in

postpartum dairy cows using ultrasound imaging and progesterone profiles. Anim.

Reprod. Sci. 22, 171~180.

75. Redmer, D.A. and Reynolds, L.P. (1996). Angiogenesis in the ovary. Rev. Reprod. 1,

182~192.

110

76. Rhodes, F.M., Fitzpatrick, L.A., Entwistle, K.W. and Death, G. (1995). Sequential

changes in ovarian follicular dynamics in Bos indicus heifer before and after nutritional

anoestrus. J. Reprod. Fertil. 104, 41~49.

77. Santos, J.E., Narciso, C.D., Rivera, F., Thatcher, WW. and Chebel, R.C. (2010). Effect

of reducing the period of follicle dominance in a timed artificial insemination protocol

on reproduction of dairy cows. J. Dairy Sci. 93, 2976~2988.

78. Sartori, R., Haughian, J.M., Shaver, R.D., Rosa, G.J. and Wiltbank, M.C. (2004).

Comparison of ovarian function and circulating steroids in estrous cycles of Holstein

heifers and lactating cows. J. Dairy Sci. 87, 905~920.

79. Savio, J.D., Thatcher, W.W., Badinga, L., de la Sota, R.L., Wolfenson, D. (1993).

Regulation of dominant follicle turnover during the oestrous cycle in cows. J. Reprod.

Fertil. 97, 197~203.

80. Schally, A.V., Arimura, A., Kastin, A.J., Matsuo, H., Baba, Y., Redding, T.W., Nair,

R.M., Debeljuk, L. and White, W.F. (1971). Gonadotropin-releasing hormone: one

polypeptide regulates secretion of luteinizing and follicle-stimulating hormones.

Science. 173, 1036~1038.

81. Senger, P.L. 2005 . Reproductive cyclicity- The follicular phase In: Senger P.L.,[Ed]

Pathways to Pregnancy and Parturition, 2nd ed.,pp. 164~187. Current Conceptions Inc.,

Pullman, Washington.

82. Shimizu, T., Jiang, J.Y., Iijima, K., Miyabayashi, K., Ogawa, Y., Sasada, H. and Sato, E.

(2003). Induction of follicular development by direct single injection of vascular

endothelial growth factor gene fragments into the ovary of miniature gilts. Biol. Reprod.

69, 1388~1393.

83. Shirasuna, K., Watanabe, S., Asahi, T., Wijayagunawardane, M.P., Sasahara, K., Jiang,

111

C., Matsui, M., Sasaki, M., Shimizu, T., Davis, J.S. and Miyamoto, A. (2008).

Prostaglandin F2 increases endothelial nitric oxide synthase in the periphery of the

bovine corpus luteum: the possible regulation of blood flow at an early stage of

luteolysis. Reproduction. 135, 527~539.

84. Siddiqui, M.A., Almamun, M. and Ginther, O.J. (2009). Blood flow in the wall of the

preovulatory follicle and its relationship to pregnancy establishment in heifers. Anim.

Reprod. Sci. 113, 287~292.

85. Siddiqui, M.A., Ferreira, J.C., Gastal, E.L., Beg, M.A., Cooper, D.A. and Ginther, O.J.

(2010). Temporal relationships of the LH surge and ovulation to echotexture and power

Doppler signals of blood flow in the wall of the preovulatory follicle in heifers. Reprod.

Fertil. Dev. 22, 1110~1117.

86. Siddiqui, M.A., Gastal, E.L., Gastal, M.O., Almamun, M., Beg, M.A. and Ginther, O.J.

(2009). Relationship of vascular perfusion of the wall of the preovulatory follicle to in

vitro fertilisation and embryo development in heifers. Reproduction. 137, 689~697.

87. Silva, L.A., Gastal, E.L., Gastal, M.O., Beg, M.A. and Ginther, O.J. (2006).

Relationship between vascularity of the preovulatory follicle and establishment of

pregnancy in mares. Anim. Reprod. 3, 339~346.

88. Sirois, J. and Fortune, J.E. (1988). Ovarian follicular dynamics during the estrous cycle

in heifers monitored by real-time ultrasonography. Biol. Reprod. 39, 308~317.

89. Smith, M.F., McIntush, E.W. and Smith, G.W. (1994). Mechanisms associated with

corpus luteum development. J. Anim. Sci. 72, 1857~1872.

90. Souza, A.H., Gümen, A., Silva, E.P., Cunha, A.P., Guenther, J.N., Peto, C.M.,

Caraviello, D.Z. and Wiltbank, M.C. (2007). Supplementation with estradiol-17β before

the last gonadotropin-releasing hormone injection of the Ovsynch protocol in lactating

112

dairy cows. J. Dairy Sci. 90, 4623~4634.

91. Sreenan, J.M., Beehan, D. and Mulvehill, P. (1975). Egg transfer in the cow: factors

affecting pregnancy and twinning rates following bilateral transfers. J. Reprod. Fertil.

44, 77~85.

92. Stevenson, J.S. (2014). Impact of reproductive technologies on dairy food production

in the dairy industry. Artificial insemination. In: Lamb, G.C. and DiLorenzo, N. [eds]

Current and Future Reproductive Technologies and World Food Production.

pp.117~119. Springer Science +Business Media, New York.

93. Stewart, R.E., Spicer, L.J., Hamilton, T.D., Keefer, B.E., Dawson, L.J., Morgan, G.L.

and Echternkamp, S.E. (1996). Levels of insulin-like growth factor (IGF) binding

proteins, luteinizing hormone and IGF-I receptors, and steroids in dominant follicles

during the first follicular wave in cattle exhibiting regular estrous cycles.

Endocrinology. 137, 2842~2850.

94. Sunderland, S.J., Crowe, M.A., Boland, M.P., Roche, J.F. and Ireland, J.J. (1994).

Selection, dominance and atresia of follicles during the oestrous cycle of heifers. J.

Reprod. Fertil. 101, 547~555.

95. Suzuki, T., Sasano, H., Takaya, H., Fukaya, T., Yajima, A. and Nagura, H. (1998).

Cyclic changes of vasculature and vascular phenotypes in normal human ovaries. Hum.

Reprod. 13, 953~959.

96. Ulbrich, S.E., Kettler, A. and Einspanier, R. (2003). Expression and localization of

estrogen receptor , estrogen receptor β and progesterone receptor in the bovine

oviduct in vivo and in vitro. J. Steroid. Biochem. Mol. Biol. 84, 279~289.

97. Vasconcelos, J.L., Sartori, R., Oliveira, H.N., Guenther, J.G. and Wiltbank, M.C.

(2001). Reduction in size of the ovulatory follicle reduces subsequent luteal size and

113

pregnancy rate. Theriogenology. 15, 307~314.

98. Wahl, M. (1985). Local chemical, neural, and humoral regulation of cerebrovascular

resistance vessels. J. Cardiovasc. Pharmacol. 7 Suppl 3, 36~46.

99. Wijayagunawardane, M.P., Miyamoto, A., Cerbito, W.A., Acosta, T.J., Takagi, M. and

Sato, K. (1998). Local distributions of oviductal estradiol, progesterone, prostaglandins,

oxytocin and endothelin-1 in the cyclic cow. Theriogenology. 49, 607~618.

100. Wiltbank, M., Lopez, H., Sartori, R., Sangsritavong, S. and Gümen, A. (2006). Changes

in reproductive physiology of lactating dairy cows due to elevated steroid metabolism.


101. Woad, K.J., Hunter, M.G., Mann, G.E., Laird, M., Hammond, A.J. and Robinson, R.S.

(2012). Fibroblast growth factor 2 is a key determinant of vascular sprouting during

bovine luteal angiogenesis. Reproduction. 143, 35~43.

102. Xiao, C.W. and Goff A.K. (1999). Hormonal regulation of oestrogen and progesterone

receptors in cultured bovine endometrial cells. J. Reprod. Fertil. 115, 101~109.

103. Xu, Z., Garverick, H.A., Smith, G.W., Smith, M.F., Hamilton, S.A. and Youngquist,

R.S. (1995). Expression of messenger ribonucleic acid encoding cytochrome P450

side-chain cleavage, cytochrome P450 17 -hydroxylase, and cytochrome P450

aromatase in bovine follicles during the first follicular wave. Endocrinology. 136.

981~989.

104. Yamagata, Y., Nakamura, Y., Sugino, N., Harada, A., Takayama, H., Kashida, S. and

Kato, H. (2002). Alterations in nitrate/nitrite and nitric oxide synthase in preovulatory

follicles in gonadotropin-primed immature rat. Endocr. J. 49, 219~226.

105. Yamashita, H., Kamada, D., Shirasuna, K., Matsui, M., Shimizu, T., Kida, K., Berisha,

B., Schams, D. and Miyamoto, A. (2008). Effect of local neutralization of basic

114

fibroblast growth factor or vascular endothelial growth factor by a specific antibody on

the development of the corpus luteum in the cow. Mol. Reprod. Dev. 75. 1449~1456.

106. Yong, E.L., Hillier, S.G., Turner, M., Baird, D.T., Ng, S.C., Bongso, A. and Ratnam,

S.S. (1994). Differential regulation of cholesterol side-chain cleavage (P450scc) and

aromatase (P450arom) enzyme mRNA expression by gonadotrophins and cyclic AMP

in human granulosa cells. J. Mol. Endocrinol. 12, 239~249.

107. Yoshioka, K., Suzuki, C., Arai, S., Iwamura, S. and Hirose H. (2001).

Gonadotropin-releasing hormone in third ventricular cerebrospinal fluid of the heifer

during the estrous cycle. Biol. Reprod. 64, 563~570.

108. Zeitoun, M.M., Rodriguez, H.F. and Randel, R.D. (1996). Effect of season on ovarian

follicular dynamics in Brahman cows. Theriogenology. 45, 1577~1581.

109. Zeleznik, A.J., Schuler, H.M. and Reichert, L.E. Jr. (1981). Gonadotropin-binding sites

in the rhesus monkey ovary: role of the vasculature in the selective distribution of

human chorionic gonadotropin to the preovulatory follicle. .Endocrinology. 109,

356~362.

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